PDBsum entry 3hlc

Transferase

PDB id

3hlc

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Contents

			Protein chain

					416 a.a. *

			Ligands

					GOL


					PG4

			Waters ×150

* Residue conservation analysis

PDB id:

3hlc

Links

Name:

Transferase

Title:

Simvastatin synthase (lovd) from aspergillus terreus, s5 mutant, unliganded

Structure:

Transesterase. Chain: a. Synonym: lovd. Engineered: yes. Mutation: yes

Source:

Aspergillus terreus. Organism_taxid: 33178. Expressed in: escherichia coli. Expression_system_taxid: 562.

Resolution:

2.00Å

R-factor:

0.181

R-free:

0.205

Authors:

M.R.Sawaya,T.O.Yeates,J.Laidman,I.Pashkov,X.Gao,Y.Tang

Key ref:

X.Gao et al. (2009). Directed evolution and structural characterization of a simvastatin synthase. Chem Biol, 16, 1064-1074. PubMed id: 19875080

Date:

27-May-09

Release date:

27-Oct-09

PROCHECK

	Headers

	References

Protein chain

Q9Y7D1 (LOVD_ASPTE) - Monacolin J acid methylbutanoyltransferase from Aspergillus terreus

Seq: Struc:					413 a.a. 416 a.a.*

Key:

PfamA domain

Secondary structure

CATH domain

* PDB and UniProt seqs differ at 8 residue positions (black crosses)



		Enzyme reactions

Enzyme class:

E.C.2.3.1.238 - monacolin J acid methylbutanoate transferase.

[IntEnz] [ExPASy] [KEGG] [BRENDA]

Reaction:

monacolin J carboxylate + (S)-2-methylbutanoyl-[2-methylbutanoate polyketide synthase] = lovastatin carboxylate + holo-[2-methylbutanoate polyketide synthase]

	Chem Biol 16:1064-1074 (2009)
PubMed id: 19875080

Directed evolution and structural characterization of a simvastatin synthase.
X.Gao, X.Xie, I.Pashkov, M.R.Sawaya, J.Laidman, W.Zhang, R.Cacho, T.O.Yeates, Y.Tang.

ABSTRACT

Enzymes from natural product biosynthetic pathways are attractive candidates for creating tailored biocatalysts to produce semisynthetic pharmaceutical compounds. LovD is an acyltransferase that converts the inactive monacolin J acid (MJA) into the cholesterol-lowering lovastatin. LovD can also synthesize the blockbuster drug simvastatin using MJA and a synthetic alpha-dimethylbutyryl thioester, albeit with suboptimal properties as a biocatalyst. Here we used directed evolution to improve the properties of LovD toward semisynthesis of simvastatin. Mutants with improved catalytic efficiency, solubility, and thermal stability were obtained, with the best mutant displaying an approximately 11-fold increase in an Escherichia coli-based biocatalytic platform. To understand the structural basis of LovD enzymology, seven X-ray crystal structures were determined, including the parent LovD, an improved mutant G5, and G5 cocrystallized with ligands. Comparisons between the structures reveal that beneficial mutations stabilize the structure of G5 in a more compact conformation that is favorable for catalysis.

Literature references that cite this PDB file's key reference

PubMed id

Reference

21351219

Y.Jiang, K.L.Morley, J.D.Schrag, and R.J.Kazlauskas (2011).
Different active-site loop orientation in serine hydrolases versus acyltransferases.

Chembiochem, 12, 768-776.

PDB code:

3ia2

20850962

C.Jäckel, and D.Hilvert (2010).
Biocatalysts by evolution.

Curr Opin Biotechnol, 21, 753-759.

20865174

C.Mura, C.M.McCrimmon, J.Vertrees, and M.R.Sawaya (2010).
An introduction to biomolecular graphics.

PLoS Comput Biol, 6, 0.

20826162

I.Campeotto, A.H.Bolt, T.A.Harman, C.Dennis, C.H.Trinh, S.E.Phillips, A.Nelson, A.R.Pearson, and A.Berry (2010).
Structural insights into substrate specificity in variants of N-acetylneuraminic Acid lyase produced by directed evolution.

J Mol Biol, 404, 56-69.

PDB codes:

2wnn 2wnq 2wnz 2wo5 2wpb 2xfw

20853106

X.Gao, P.Wang, and Y.Tang (2010).
Engineered polyketide biosynthesis and biocatalysis in Escherichia coli.

Appl Microbiol Biotechnol, 88, 1233-1242.

The most recent references are shown first. Citation data come partly from CiteXplore and partly from an automated harvesting procedure. Note that this is likely to be only a partial list as not all journals are covered by either method. However, we are continually building up the citation data so more and more references will be included with time. Where a reference describes a PDB structure, the PDB code is shown on the right.